Re: Seeking H&E stains procedure for Ground Plastic section

From:

Bryan Llewellyn

Bernard,

I never worked with plastic sections at 40-100 micrometres as
you are. I used a butyl/methyl methacrylate mixture and cut one
micrometre renal biopsies, so the protocol below may or may not be useful to
you. It worked quite well for what I was doing.

1. Remove plastic with acetone for one hour
at RT. This may not be feasable for 40 micron sections but is worth a
try.

2. Wash with water to ensure the sections
are wetted.

3. Place in preheated Mayer's or Cole's
hemalum for an hour or longer. Change the time to get the desired
depth of staining.

4. Wash with water and blue. I used
0.5% aqueous sodium acetate.

5. Wash with water.

6. Counterstain with eosin Y ws.
Adjust temperature, time and concentration to get what you want.

7. Rinse with water to remove excess
etc.

8. Dehydrate, clear and mount as
usual.

Both Mayer's (Langeron's) and Cole's hemalums are
progressive, so require no differentiation. Mayer's is more selective for
nuclei, Cole's staining is similar to a differentiated regressive stain.
Formulae are on StainsFile.

My Pathologist is adamant in me, finding a H&E procedure
that will work, to stain some plastic processed (Methyl Methacrylate) pig
& rabbit eye implants. I mentioned to him about other stains out
there, that work similarly to the H&E and stains
better. However, he insists that the H&E will give him a
familiar and better contrast.

The one method I have, worked
(acceptable to him) for another plastic procedure( A Butyl Methyl
Methacrylate), but not this one. The stain came out to light.
I even restained the slides following the same procedure again. And it
still turned out light.